Process Biochemistry 26 (1991) 101-107

Kinetics of Methane Production from Olive Mill

Wastewater

A. Martin, R. Borja, b I. Garcia a & J.A. Fiestas

"Department of Chemical Engineering, Faculty of Sciences, Avda San Alberto Magno s/n, E-14004 C6rdoba,
Spain.
OInstitute of Fat and its Derivatives, Avda Padre Garcia Tejero 4, E-41012 Sevilla, Spain

(Received 26 September 1990; revised version received 10 March 1991; accepted 23 March 1991)

A kinetic study on the anaerobic digestion o f olive mill wastewater

( O M W ) was carried out in a semicontinuous, well-stirred fermenter
with microorganisms immobilized on clayey supports.
Substrate concentrations equal to or higher than 3.5 g COD/litre
within the reactor resulted in a methane production rate that was first-
order with respect to biodegradable substrate concentration. The
apparent rate constant was proportional to the VSS concentration and
varied slightly with the nature o f the support.
The yield coefficient, Yws, was 0"26 litres C H 4 S T P / g COD
irrespective o f the support used.

INTRODUCTION transforming most of the organic substances present

in the waste into methane, (c) sludge formation is
The manufacturing process of olive oil usually minimal, (d) nutrient demands are very low, and (e)
yields an oily phase (20 %), a solid residue (30 %) unpleasant odours are avoided.
and an aqueous phase (50%) formed from the The low growth rate of anaerobic micro-
water content of the fruit. Such water, when organisms has encouraged the development of
combined with that used in washing and processing
the olives, makes up the so-called 'olive mill Table 1. Overall Composition of Olive Mill Wastewater
wastewater' (OMW) and also contains soft tissues
from olive pulp and a very stable oil emulsion. Batch Continuous
Table 1 summarizes the overall composition of a Parameter process process
typical OMW.
pH 4"5-5 4-7-5.2
The high polluting power and large volumes of COD 120-130 45-60
OMW produced along the Mediterranean basin BOD 90-100 35-48
pose large-scale environmental problems. 1 Among SS 1 9
TS 120 60
the feasible alternatives for the purification and MS 15 5
exploitation of OMW, anaerobic fermentation has VS 105 55
a number of advantages, namely :1, 2 (a) it demands Fat 0.5-1-0 3-10
little energy, (b) anaerobic bacteria are capable of
All concentrations are expressed in g/litre.
SS, TS, MS, VS = suspended, total, mineral, volatile solids.
Corresponding author: Dr A. Martin. Telephone: (957)218624 COD = chemical oxygen demand.
ext. 217; Fax: (957)218606. BOD = biological oxygen demand.

Process Biochemistry 0032-9592/91/$3.50 © 1991 Elsevier Science Publishers Ltd, England.

102 A. Marffn, R. Borja, L Garcia, J.A. Fiestas

various techniques for their immobilization within
bioreactors in order to avoid loss in the effluents
stream and hence diminished process rates. Among
the bioreactors more commonly used for this
purpose are fluidized bed reactors, where bacteria
colonize particles of some materials, thereby in-
creasing the surface available for bacterial growth.
These reactors also cope with the greatest volu-
metric loads (larger than 50 k g C O D / m 3 day),
which makes them suitable for the treatment of
wastewater with high organic loads (e.g.
OMW). 3
The significance of the nature of the support
material to be colonized by the bacteria has been
emphasized by several authors, 4' 5 since it influences
the production and composition of the biogas and
the proportions in which the different types of
microorganism develop in the overall anaerobic
biomass. 6
The aim of this work was to carry out a kinetic
study on the anaerobic purification of OMW in
bioreactors containing immobilized microorgan-
isms and to determine the yield coefficient, Yws,
which is a measure of the extent of transformation
of the biodegradable substrate into methane.
MATERIALS AND M E T H O D S
Equipment
A well-stirred anaerobic fermenter with a working
volume of 1.12 litres and furnished with a settling
mechanism intended to avoid loss of the solids
acting as supports for the microorganisms was
employed. The reactor was fed and liquid effluents
were removed several times a day. The gas was
evacuated in a continuous fashion; the volume of
methane produced being measured after removing
CO 2 by adsorption into NaOH.
Supports
The supports used to immobilize the bacterial
biomass were of clayey nature, namely montmor-
illonite and sepiolite (Pansil), which were selected
on account of their favourable kinetic behaviour.
Their features are described in detail elsewhere. 7
Inoeulum
Digesters were inoculated with sludges from an
OMW pool after appropriate dilution and neutral-
ization. The inoculum composition is given in Table
2.
Olive mill wastewater
The features of the OMW used are summarized in
Table 3.
Experimental procedure
Two anaerobic digestion units contained alter-
natively montmorillonite or sepiolite, as support.
To each was initially added 750 ml distilled water,
250 ml inoculum and 10 g support. Prior to semi-
continuous experiments, the digesters were con-
ditioned by successive additions of increasing
concentrations of the OMW.
This preliminary stage was followed by four
series of semicontinuous experiments performed
with OMW contents of 10 %, 20 %, 40 % and 80 %
as influent with 25, 35, 40, 50, 60 and 100 ml/day
feeding flow-rates, which are equivalent to 44.8,

Table 2. Composition of the Biomass Used as Inoculum

TS VS MS TSS VSS MSS pH

16-3 12.9 3.4 13"0 10.4 2"6 7"0

All concentrations are expressed in g/litre.

TSS, VSS, MSS = total suspended, volatile suspended, mineral suspended solids.

Table 3. Features of the OMW Used

Total
Volatile polyphenols
acidity Alkalinity Nitrogen (tannic
pH COD TS MS VS TSS MSS VSS (HAcO) (CaCOz) (NH~) acid)

5"0 50-0 47'0 12"0 35"0 11"9 2.0 9-9 0"11 0'50 0'05 0'47

All concentrations are expressed in g/litre.

 Kinetics o f methane production f r o m olive mill wastewater 103

32-0, 28.0, 22-4, 18-6 and 11.2 days of hydraulic Table 6. Variation of the Effluent pH with the O MW Flow-rate
(ml/day) in the Digester Containing Montmorillonite as
retention time, respectively. Support
Once steady-state conditions were approached
and at each feeding flow-rate the volume of methane Flow-rate
produced and the pH and COD of the different COD
(inlet) 25 35 40 50 60 100
effluents were determined. After the experiments
with a constant COD flow to the digester were 5"0 7-4 7"5 7"4 7"4 7"5 7'4
completed the VSS within the reactor was de- 10'0 7"5 7"4 7"4 7"3 7"3 7"4
termined. 20'0 7-4 7"3 7"3 7-2 7"2 7"2
40"0 7-4 7-4 7"3 7-3 7'3 7'4

Chemical analyses
Analyses were performed according to the guide- Table 7. Variation of the Effluent pH with the O MW Flow-rate
lines of the Standard Methods for the Examination (ml/day) in the Digester Containing Sepiolite as Support
of Water and Wastewater. 8
Flow-rate
COD
(inlet) 25 35 40 50 60 100
RESULTS
5.0 7.5 7-4 7-3 7.3 7.2 7.3
The volumes of methane produced during periods 10-0 7.5 7-6 7.4 7.4 7.3 7.3
20-0 7-4 7.3 7.3 7.3 7'2 7'3
of 24 h, flow-rates and COD values are listed in 40.0 7.4 7'3 7'3 7.2 7'3 7"3
Tables 4 and 5, while Tables 6 and 7 list the
variations of the pH at the digester outlet.
The substrate concentrations measured at the Table 8. Variation of the COD (g/litre) within the Reactor
Containing Montmorillonite as Support with the Flow-rate
digester outlet are given in Tables 8 and 9. As a (ml/day)
fraction of this was not biodegradable, it was
estimated in order to perform the subsequent Flow-rate
COD
calculations; this was accomplished by a graphical (inlet) 25 35 40 50 60 100
procedure involving extrapolation of the substrate
concentration at an infinite residence time (Figure 5'0 1'0 1"1 1"3 1-5 1"6 1-9
1). 10.0 1"1 1-2 1-3 1-5 1"7 2-2
20"0 1"3 1"5 1-7 1"9 2'3 2"6
40-0 1"6 1"8 2-0 2-4 2"9 3-5
Table 4. Volume of Methane STP (ml) Obtained at Different
O M W Flow-rates (ml/day) Using the Digester Containing
Montmorillonite as Support
Table 9. Variation of the COD (g/litre) within the Reactor
Flow-rate Containing Sepiolite as Support with the Flow-rate (ml/day)
COD
(inlet) 25 35 40 50 60 100 Flow-rate
COD
5"0 27 37 41 49 57 83 (inlet) 25 35 40 50 60 100
10.0 52 70 79 93 110 168
20.0 110 150 168 204 240 380 5'0 1-0 1.1 1.2 1.4 1.6 2"0
40.0 285 378 424 524 616 976 10.0 1-1 1.2 1.4 1.6 1.7 2'0
20.0 1.2 1.4 1.5 1.7 2.1 2.5
40.0 1-5 1'7 1.9 2.2 2'5 3.0
Table 5. Volume of Methane STP (ml) Obtained at Different
OMW Flow-rates (ml/day) Using the Digester Containing
Sepiolite as Support Table 10. Variation of the Biomass Concentration, VSS
(g/litre), within each Digester as a Function of the Feed COD
Flow-rate (g/litre)
COD
(inlet) 25 35 40 50 60 100 COD (inlet)

5.0 27 36 40 49 56 80 Support 5"0 10"0 20"0 40"0

10.0 61 81 90 105 125 175
20-0 119 160 178 215 252 382 Montmorillonite 3"0 3.1 3'6 3.9
40-0 290 392 440 536 629 1000 Sepiolite 4.3 4-4 4.6 5'0
104 A. Mart(n, R. Borja, L Garc(a, J.A. Fiestas
-S
sumption and the specific rate of microbial growth,
# (day-a), are related through 9
(-- G) Yx/~ = #X (4)
where Yx/~ is the growth yield constant (g cell
mass/g COD) and X is the cell mass concentration
(g/litre).
For a system whose specific growth rate conforms
to the M o n o d equation:
rG = [( YwsXflm)/Yx/j[S/(K~ + S)] (5)
At low substrate concentrations, K s >> S, so eqn
~, lO% (5) simplifies to
+ 20%
C) 40%
FG = ( Yp/s tim/Yx/s Ks)XS (6)
[] 80% For narrow time intervals, as anaerobic micro-
organisms grow very slowly:
0 0.02 0.04 0.06 0.08
1/ @(day-1 ) r a ~- K S (7)
Fig. 1. Estimation o f the non-biodegradable matter contained where
in the processed residue.
K = ( Yp/s~/m/Yx/s K s ) X (8)
Although, strictly speaking, the experimental set-
Finally, Table 10 lists the biomass concentrations,
up is not continuous and the working conditions
expressed as VSS, in the bulk reactor found after
are not strictly steady-state conditions, the slow
the experiments performed using 5, 10, 20 and 40
evolution of the system concerned allows one to
g/litre as entry COD.
assume that these two conditions are roughly met.
Figures 2 and 3 shows the variation of the rate of
gas production as a function of the biodegradable
DISCUSSION substrate concentration. As can be seen, the paired
For a continuous well-stirred fermenter working
F rG(I OH4 /I.day)
under steady-state conditions

(--G) = ( S o - S ) / O (1)
where ( - r s ) is the substrate uptake rate (g
80%
COD/litre day), S o and S are the substrate
concentrations (g COD/litre) at the fermenter inlet /
/
and outlet, and 0 is the retention time (day). 0.6 //D
If the volume o f gas produced, VO<~CH,Srp>, is //
assumed to be proportional to the a m o u n t of /
substrate consumed, then
V~ = Yws(So - S)q (2)
where Yw~ is the yield coefficient (litres CH 4 STP/g
COD), and q is the volumetric feeding flow-rate
(litres/day).
This equation allows one to relate the rate of
substrate uptake to that of gas production, r~,
o o.s i 1.s 2 2.s
through the following expression: S (g COD/I)

rG = Yws( -- G) (3) Fig. 2. Variation of the gas production rate as a function of the
biodegradable substrate concentration. Support: montmor-
On the other hand, the rate of substrate con- illonite.
 Kinetics of methane production from olive mill wastewater 105
'~ ~ 10% / --r G (CH4 /I.day)
--+-
-~ 40%
20%
/
[] /
l
0.8 -_E~ 80% /
0.8

0.6
[]
0.6 Q
[]

0.4
[] ©

Q
0.2
~ J
0.2 ~J ~- lo%
--¢- 20%
40%
0
0 0.5 1 1.5 2 2.5 3 80%

S (g COD/I) 0 I I I I I I I II I I J I I I I I

0.1 1 10
Fig. 3. Variation of the gas production rate as a function of the S (g COD/I)
biodegradable substrate concentration. Support: sepiolite.
Fig. 5. Plot of eqn (8) in logarithmic coordinates. Support:
values (rQ, S) can be fitted to a straight line of zero sepiolite.
intercept which, in principle, suggests that eqn (7) is
valid for the process studied. In order to check this two reactors used. The straight line obtained and its
hypothesis we used a twofold procedure. First, by slope (close to 1) confirm the validity of the assayed
converting eqn (7) to its logarithmic form one has: model.
On the other hand, if X is assumed to remain
In r~ = In K + In S (9) constant t h r o u g h o u t a series of experiments, then
according to which, the plot of In r G against In S eqn (5) can be written as:
should be a straight line of unity slope and intercept r G = KI[S/(K s + S)] (10)
equal to In K. Figures 4 and 5 show this plot for the
where
r G (I CH 4 /I.day)
g 1 = XYp/s~lmlYx/s (11)
F
Linearization of eqn (10) yields:
1/r~ = ( 1 / K 1 ) + ( K s / g l ) ( 1 / S ) (12)

Figure 6 shows the aforesaid plot for one of the

cases studied; the nearly zero intercept of the line
0.1 again confirms the validity of the assayed model.
By fitting the (rG, S) data pairs to a straight line
the constant defined in eqn (8) is obtained, the
values of which are listed in Table 11.
In order to check the goodness of fit the
0.01 experimental reaction rate was plotted against its
J ~- 10% theoretical counterpart (Figure 7), which was
--f-- 20%
calculated from the kinetic constants listed in Table
-0- 40%
11. As can be seen, the experimental results were
80%
reproduced with errors equal to or less than 10 % in
0.001 i i ~ ~ ~ ~J L I I I I I I 1
0.1 10 every case.
S (g COD/I) According to eqn (8), K m u s t be proportional to
Fig. 4. Plot of eqn (8) in logarithmic coordinates. Support: the microorganism concentration, which is con-
montmorillonite. firmed by Figure 8, a plot of this variable against
106 A. Mart(n, R. Borja, I. Garc(a, J.A. Fiestas
1/r G (I.day/I CH 4) Table 11. Variation of K (litres CH 4 STP/day g COD) in Eqn

41
(8) with the Influent Content for the Two Types of Support
~- 10% Assayed.
~--- 20%
A OMW
-~:~ 4OVo
/ content in Support
80%
the influent
/ (%) Montmorillonite Sepiolite
//
/
///\
30 10 0'05 0-04
20 0"09 0"10
/
40 0"16 0"18
80 0"30 0-39
20
// [ K (I CH4/day * g COD) /

Montrnorillonite
Pansil
t / / / / Z j-~ +
0.3

0 0.5 1 1.5 2
1/S (I/g COD)
Fig. 6. Lineweaver-Burk plot for the digester with sepiolite as o.2 i
support.

- r G (experimental)
0.1

O.
•,oo, V1 ,/
4
jd. lOO, o
0
2.5 3.5 4.5 5.5
O. x (VSS/0
Fig. 8. Variation of the apparent rate constant as a function of
volatile suspended solids (VSS).

r VG/0 (I/day)

j/
0.4

E~ 10% 1.0
0.2
-~ 20%
Q 40%
//
CI 80% 0.8 /
0 ~ - ~ - L L
/
0 0.2 0.4 0.6 0.8
r G (calculated) 0.6
Fig. 7. Comparison between the experimental rates of gas
production and that predicted by eqn (8). /
0.4
///
the concentration of suspended volatile solids. The [ L!//
non-zero intercept, contrary to the above equation, 10%
~o
0.2 ÷ 2O~o
may be a result of the form in which the
© 40%
microorganism concentration is expressed. 10 In fact,
80%
the experimental procedure used does not dis-
L
tinguish between active and inactive microorgan- 0 1 2 3
isms; in addition, non-viable organic components (So - S) * q (g COD/day)
may be assayed in the process. Fig. 9. Determination of the yield coefficient (Yp/s).
 Kinetics o f methane production from olive mill wastewater
Finally, the experimental data listed in Tables 4,
5, 8 and 9 were used to determine the yield
coefficient for each of the fermenters and supports.
By plotting eqn (2) in the form ( V J t ) v s [(S0 - S)q]
(Figure ,9), we obtained a virtually constant value
over the substrate concentration range assayed, The
average value was 0"26 litres CH 4 STP/g COD and
was independent of the type of support used; in
addition, it is consistent with the report of Chen and
Hashimoto) This value, together with those of the
overall and biodegradable COD, allows the es-
timation of the energy one can obtain from the
anaerobic purification process.
CONCLUSIONS
The anaerobic digestion of olive mill wastewater
conforms to first-order kinetics at substrate concen-
trations equal to or lower than 3.5 g COD/litre.
The apparent rate constant of the process, K, is
proportional to the microorganism concentration
and varies slightly with the nature of the support.
The experimental kinetic constants reproduce the
reaction rate values with errors not larger than 10 %
in any case.
The yield coefficient, Yws is 0.26 litres CH 4 STP/g
COD and is independent of the type of support
used.
9 E P B 26
107
ACKNOWLEDGEMENT
The authors wish to thank the Consejeria de
Educaci6n y Ciencia de la Junta de Andaluc{a for
financial support granted for the realization of this
work.
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